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文献和实验. Directions for preparation of the DNaseI column are below in Appendix . Day 1 Extraction of Actin from Chicken Brain (large scale prep from tissue) 1. Homogenize tissue (fresh or frozen) in 1X Extraction Buffer without detergents at a ratio of 100
Isolation of Actin and Myosin Filaments
7.0 0.001 M EDTA (Ethylene diamine tetra acetic acid) Blender Preparative centrifuge Materials for TEM fixation, embedding and observation Procedure 7 Obtain fresh chicken gizzards 8 and dissect the lateral muscles
LABELING MUSCLE ACTIN WITH IATR
1.Centrifuge in a 50Ti (1 hr,40,000 rpm)or 42.2Ti (30 min,25,000 rpm)at 4℃. 2.Measure concentration and dye/protein molar ratio.Dilute 1:40 with the dialysis buffer and read the OD at 555 nm. D/P = {OD555 x 41 / 60,000} / {(mg/ml)/ 43,000
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